Description
The 11F2 monoclonal antibody specifically reacts with a framework epitope of the γ/δ TCR. The γ/δ TCR is a heterodimeric glycoprotein that is noncovalently associated with the CD3 antigen complex. The γ and δ TCR chains are composed of constant and variable regions, each encoded by distinct gene segments. The γ chain forms either disulfide-linked or non-disulfide-linked heterodimers with the δ-subunit. TCR γ/δ is present on a minor subset of T lymphocytes in peripheral blood, thymus, spleen, and lymph node. TCR γ/δ-positive T lymphocytes comprise 1% to 9% of normal peripheral blood lymphocytes and less than 2% of normal thymocytes. The 11F2 antibody is mitogenic for γ/δ-TCR-bearing T lymphocytes.
Bead-based compensation or unmixing controls, such as BD® CompBeads or BD™ SpectraComp™, can be used as surrogates to assess fluorescence spillover when bound to fluorochrome-conjugated antibodies. Although these beads have spectral properties similar to cells, variations in spectral emission may occur, resulting in differing spillover values compared to biological controls. Therefore, it is considered best practice to compare the spillover obtained from cells and bead-based controls when using BD® CompBeads or BD™ SpectraComp™ for the first time, to ensure they are appropriate for the intended application.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions that minimize unconjugated dye and antibody.
1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
2. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
3. For U.S. patents that may apply, see bd.com/patents.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
6. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
7. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
9. An isotype control should be used at the same concentration as the antibody of interest.
10. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
11. Alexa Fluor™ is a trademark of Life Technologies Corporation.
12. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
Specifications

General

SourceHuman B7-2/CD86, amino acid Leu26-Pro247 (accession # NP_787058.4) with a C-terminal 6x His tag, was expressed in 293E cells.
Molecular MassThe 228 amino acid recombinant protein has a predicted molecular mass of approximately 26.21 kD. The DTT-reduced and non-reduced protein migrates at approximately 52 kD by SDS-PAGE. The predicted N-terminal amino acid is Leu.
Purity> 95%, as determined by Coomassie stained SDS-PAGE.
Formulation0.22 µm filtered protein solution is in 1x PBS.
Endotoxin LevelLess than 0.1 EU per µg protein as determined by the LAL method.
Concentration10 and 25 µg sizes are bottled at 200 µg/mL. 100 µg size and larger sizes are lot-specific and bottled at the concentration indicated on the vial. To obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.
Storage & HandlingUnopened vial can be stored between 2°C and 8°C for up to 2 weeks, at -20°C for up to six months, or at -70°C or colder until the expiration date. For maximum results, quick spin vial prior to opening. The protein can be aliquoted and stored at -20°C or colder. Stock solutions can also be prepared at 50 - 100 µg/mL in appropriate sterile buffer, carrier protein such as 0.2 - 1% BSA or HSA can be added when preparing the stock solution. Aliquots can be stored between 2°C and 8°C for up to one week and stored at -20°C or colder for up to 3 months. Avoid repeated freeze/thaw cycles.
ActivityWhen human B7-2/CD86 is immobilized at 0.5 μg/ml (100 μl/well) using a anti-His antibody (Cat. No. 652501), biotinylated human CTLA-4 binds with EC50 of 10 - 40 ng/ml in a functional ELISA.
ApplicationBioassay
Application NotesBioLegend carrier-free recombinant proteins provided in liquid format are shipped on blue-ice. Our comparison testing data indicates that when handled and stored as recommended, the liquid format has equal or better stability and shelf-life compared to commercially available lyophilized proteins after reconstitution. Our liquid proteins are verified in-house to maintain activity after shipping on blue ice and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at [email protected].
StructureImmunoglobulin family
DistributionActivated antigen presenting cells, including B cells, dendritic cells and macrophages
FunctionB7-2 is a costimulatory molecule that is required for T cell proliferation and IL-2 production.
InteractionLeukocytes, endothelial cells
Ligand/ReceptorCD28, CTLA4
BioactivityMeasured by the ability of immobilized protein to bind biotinylated human CTLA-4
Biology AreaCell Biology, Costimulatory Molecules, Immunology, Neuroscience, Neuroscience Cell Markers
Molecular FamilyCD Molecules, Immune Checkpoint Receptors, Soluble Receptors
Gene ID942
UniProtView information about B7-2/CD86 on UniProt.org
Regulatory StatusRUO
Other NamesCD86, B7-2, T lymphocyte activation antigen CD86, Activation B7-2 antigen, CD28 antigen ligand 2, CD28LG2B7-2 antigen, B70, BU63, CTLA-4 counter-receptor B7.2, FUN-1